GSM3746498: Nup93 ChIPSeq rep2; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
NUP93
Cell type
Cell type Class
Digestive tract
Cell type
DLD-1
Primary Tissue
Colon
Tissue Diagnosis
Adenocarcinoma
Attributes by original data submitter
Sample
source_name
Nup93_ChIPSeq
cell line
DLD-1
cell type
Colorectal cancer cells
chip antibody
Anti-Nup93 antibody (sc-292099)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Crosslinking 1% paraformaldehyde for 10 min, nuclear purification with 0.5% NP40, DNA solubilization and sonication in 1% SDS using Covaris sonicator, followed by Immunoprecipitation with following antibodies 1.Anti-Nup93 antibody (sc-292099)- 2µg/ 100µg of chromatin, 2. Normal Rabbit IgG (Millipore 12-370). Arpoximately 20-30 ng of DNA was used for library preparation. ChIP-Seq libraries for sequencing were constructed according to the NEXTflex™ ChIP-Seq library protocol outlined in NEXTflex™ ChIP-Seq Kit - 5143-01. Briefly, DNA was subjected to a series of enzymatic reactions that repair frayed ends, phosphorylate the fragments, add a single nucleotide A overhang and ligate adaptors (NEXTflex ChIP Barcodes-48 kit). The libraries are enriched using PCR (5 cycles), fragments were size selected using a 2 % Low melting agarose gel and purified using MinElute Gel Extraction Kit (QIAGEN). The libraries were further enriched using PCR (14 cycles), post PCR cleanup was performed using Agencourt AMPURE XP beads (Beckman Coulter #A63881). The prepared libraries were quantified using Qubit fluorometer and validated for quality by running an aliquot on High Sensitivity Bioanalyzer Chip (Agilent).